WO2009042080A1

WO2009042080A1 - Rapid assays for determining mhc-correlated profiles for matching of potential mating partners

Info

Publication number: WO2009042080A1
Application number: PCT/US2008/010936
Authority: WO
Inventors: August Hammerli; Manuel Kaegi; Dominic Senn; Hanna Hammerli
Original assignee: Basisnote Ag
Priority date: 2007-09-21
Filing date: 2008-09-19
Publication date: 2009-04-02

Abstract

The present invention provides a user friendly test kit for determining an individual's Major Histocompatibilty profile, transmitting such information in digital form, and using that profile in an internet-based format to determine optimal mating matches between individuals, preferably in conjunction with other matching parameters.

Description

Rapid Assays for Determining MHC-Correlated Profiles for Matching of

Potential Mating Partners

FIELD OF THE INVENTION

The present invention relates to the rapid, non-diagnostic, non-therapeutic assay for MHC or MHC-correlated profiles and to the matching of such profiles as a means of determining optimal mating compatibility between individuals. Particularly, the invention is applicable for the encrypted and anonymous matching of optimal mating partners in a digital format as may be applicable in such fields as internet-based dating services.

BACKGROUND OF THE INVENTION

The major histocompatibility (MHC) gene complex encodes cell-surface glycoproteins (class I and Il molecules) that bind peptides and present them to T lymphocytes [1]. While the MHC complex has been detected in the field of medicine, specifically in transplantation medicine, the high degree of heterozygosity in this gene complex found in natural populations of most species is most likely promoted by non-disease-based selection such as mating preferences [2]. Evolutionary biologists have been investigating MHC-dependent mating preferences in vertebrates [3] and particularly in humans [4]. Evidence is accumulating that in many species, MHC genes influence reproductive behaviour and body odour, generating MHC-correlated olfactory cues for potential mating partners [3, 5]. The high diversity of MHC genes may be responsible for the distinctive scent of individuals, which is analogous to a signature or fingerprint [6]. Such MHC-correlated odour fingerprints consist of various volatile odour compounds and MHC peptide ligands [7].

Empirically, in the three spined stickleback fish, evolutionarily conserved structural features of MHC peptide ligands were used to evaluate MHC diversity of prospective mating partners through MHC-correlated olfactory cues [8]. In human behaviour, odour cues provide information regarding the degree of relatedness and may thus affect kin preferences and mate choice [9]. For example, it has been shown that humans can influence the hormonal balance of conspecifics through chemosignals from sweat [10]. Evidence is accumulating that olfactory cues on MHC status of individuals are also possibly enhanced through the use of perfumes. Individual preferences for fragrances seem to be determined in some way by amplifying one's own body odour that reveals a person's immunogenetics [11]. Odour cues that reveal the immunogenetic status of individuals are highly resolved. Women for example have been shown to be able to differentiate a single allele difference among male odour donors with different MHC genotypes [12]. MHC-dependent mate preferences through odour cues may even influence the psychology of sexual attraction and the number of extra- pair sexual partners in relationships [13]. Evolutionary biologist assume that the optimal MHC complement for a partner should include those alleles that provide resistance against the parasites in the current environment [14]. Theoreticians have predicted two opposing forces, namely parasite resistance and inbreeding avoidance, to result in an optimal number of genes at intermediate individual MHC diversity [15, 16]. This prediction is now supported both by experimental data [17] and a population genetic survey [18] with the three spined stickleback.

MHC linked odour cues appear at the periphery via different pathways. Soluble MHC molecules for example are known that carry allele-specific odoriferous molecules from the blood via the kidneys into the urine [5] and such molecules have been identified in the urine proteome [19], where carboxylic acids are most likely to be the odour-components linked to MHC [20]. More and more data is becoming available on odour components not only in urine but also sweat and saliva and many of these components seem to be MHC determined [6]. Volatile odorant components have been experimentally shown to activate neurons in the vomeronasal organ [21]. And among those, small peptides that serve as ligands for MHC molecules function as sensory stimuli [22]. Fish, rodents and humans have been shown to be able to differentiate particular MHC profiles by smell [20] and such differences could in principle even be detected with artificial screening technology (electronic nose) [23] [24]. Data on these ligands is rapidly accumulating [25] together with their specific binding properties to MHC molecules [26]. The following paragraph reviews the currently available methods for the screening of MHC profiles.

The screening for MHC genes or MHC correlated profiles is generally directed towards medical, diagnostic purposes, the methodologies focusing on sequencing of the MHC genes or the use of genetic markers for which often blood samples have to be collected. Invasive sampling procedures and time consuming methodologies may be among the reasons why odour correlated MHC profiles have not yet been fully exploited in the life-style market. In the medical field, however, several methodologies for screening MHC profiles other than sequencing and genotyping exist. Medical research has explored the possibility for enzyme linked immunosorbent assays (ELISA) [27] to screen not only blood but also non-invasive urine, saliva and sweat samples. Typically, in such ELISA protocols, anti- monomorphic MHC class I or class Il monoclonal antibodies are attached to solid substrate beads, and promote the capture of soluble MHC molecules from serum, plasma or other human body fluids [28]. The detected MHC linked proteins were typically between 23 and 45 kD in molecular weight [29] and the amount of soluble MHC molecules in saliva, for example, ranged from 9 ng/ml to 70 ng/ml [30]. However, despite the diversity of available ELISA protocols for MHC screening that have been established [31, 32], its success for medical diagnostic purposes has been limited. Either the accuracy of the assays was insufficient for medical standards compared to sequencing and/or other serological analysis, or the diagnostic power for diseases with a demand for non-invasive screening was too low (e.g. for rheumatoid arthritis) [31]. However, ELISA based protocols, in general, seem to be the most promising candidate methods for the manufacture of a rapid test in a lateral flow setting [33, 34]. Interestingly, in the context of ELISA, it has become clear that MHC molecules themselves are not completely broken down in the body but are being excreted and can themselves act as recognition cues [29]. The aim of the present invention is to manufacture a rapid assay that could be used by individuals at home. The assay results are such that they can be added to a personal profile on an Internet platform.

Internet communities such as online dating services explore personal profiles including images and other personal attributes such as body shape [35, 36] and facial characteristics [37] to determine attractiveness [38] and to bring together individuals with similar backgrounds [39]. The market volume in this field for Europe and the US alone, is between 2.5 and, 5 million paying singles using Internet dating platforms and an industry generating a sales volume of approximately 1 ,200 million USD per year. US patent application 2005/0112684 A1 proposes a method of matching human beings, which comprises collecting genetic sample material from individuals and subsequently determining the individuals¹ MHC profiles in a genetic laboratory. Similarly, US 2007/069889 A1 and US 2007/069901 disclose matching services which may include the determination of MHC profiles. These methods, however, require that the individuals give their genetic material to an institution where it is analyzed. It is likely that the users or participants will be reluctant to give their genetic material and have it analyzed at a location where it is beyond their control.

SUMMARY OF THE INVENTION

Accordingly, it is one of the objects of the present invention to provide a user-friendly test kit for the rapid determination of an individual's Major Histocompatibility Complex (MHC) profile, comprising: at least one solid substrate in contact with a proximal sample application zone, said substrate having N conjugation zones and N distal detection zones, said conjugation zones respectively containing N labelled primary binding reagents capable of binding with an analyte to form an analyte-primary binding reagent conjugate; said distal detection zones respectively having immobilized thereunto, N unlabeled secondary binding reagents capable of binding to a migrating analyte-labeled primary binding reagent conjugate to determine the presence or absence of N analytes which are indicative of defined N-MHC antigens, wherein N is an integer greater than 3.

Another object of the invention is to provide a user-friendly test kit for the rapid determination of an individual's Major Histocompatibility Complex (MHC) profile, comprising: at least one solid substrate in contact with a proximal sample application zone, said substrate having N conjugation zones and N distal detection zones, said conjugation zones respectively containing N labelled primary binding reagents capable of binding with an analyte to form an analyte-primary binding reagent conjugate; said distal detection zone respectively having immobilized thereunto, N unlabeled secondary binding reagents capable of binding to the respective labelled primary binding reagent, but not to a migrating analyte-labeled primary binding reagent conjugate to determine the presence or absence of N analytes which are indicative of defined N- MHC antigens, wherein N is an integer greater than 3.

A further object of the invention is to provide a user-friendly test kit for the rapid determination of an individual's Major Histocompatibility Complex (MHC) profile, comprising: at least one solid substrate in contact with a proximal sample application zone, said substrate having N release zones and N distal detection zones, said release zones respectively containing N labelled analytes; said distal detection zones respectively having immobilized thereunto, N unlabeled binding reagents capable of binding to a migrating unlabelled analyte or to a migrating labelled analyte to determine the presence or absence of N analytes which are indicative of defined N-MHC antigens, wherein N is an integer greater than 3.

Another object of the invention relates to a method of matching individuals, comprising the following steps:

(a) obtaining a plurality of personal profiles from a plurality of individuals and storing said personal profiles in a database on a computer.

(b) providing a plurality of test kits to said plurality of individuals, wherein each test kit comprises a unique identifier;

(c) obtaining a plurality of encrypted MHC profiles and the respective unique identifiers from said plurality of individuals;

(d) storing said encrypted MHC profiles and the respective unique identifier in a database on a computer, wherein each encrypted MHC profile and its unique identifier is allocated to one of said personal profiles;

(e) decrypting said encrypted MHC profiles using a key related to said unique identifier, and a computer, and storing the decrypted MHC profiles in a database on a computer;

(f) calculating the pair wise distances in decrypted MHC profiles between all individuals for which decrypted MHC profiles have been stored in the database;

(g) determining the quality of match between said individuals from the pair wise distances calculated in step (f).

Another object of the invention relates to a method for the rapid determination of an encrypted MHC profile of an individual in which a test kit according to the invention is contacted with a body liquid sample containing the analyte, such that the sample permeates by capillary action from a sample application zone through the solid substrate via N conjugation zones into N detection zones, and N labelled primary binding reagents migrate with the analyte from the respective conjugation zones to the respective detection zones containing immobilized N unlabelled secondary binding reagents, the presence of analytes in the sample being determined by observing the extent to which the labelled reagents become bound in the detection zones.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the invention as claimed. Other objects and features of the invention will become apparent from the following detailed description considered in conjunction with the accompanying drawings and tables. The accompanying drawings are included solely for purposes of illustration and not as a definition of the limits of the invention. Also, the drawings are not drawn to scale, and are merely conceptual in disclosing the preferred embodiments of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 illustrates a process flow-chart of the processes according to one embodiment of the present invention.

Figure 2a illustrates one embodiment of a lateral flow-through assay device for rapid assay to detect the encrypted MHC profile of a user.

Figure 2b illustrates how the result from such a flow-through assay device can be added to a personal profile in an online electronic form together with a unique identifier.

Figure 3 is a process chart of matching and dating services according to one embodiment of the present invention showing in chronological order, the preferred steps usable to provide a user with a list of potential dating partners that match said user's MHC profile.

Figure 4 is an object-oriented view of matching and dating services according to one embodiment of the present invention showing the objects, methods and database tables for providing a user with a list of potential dating partners that match said user's MHC profile. Figure 5 is a chart illustrating the the probability P₁ with which a test field shows a positive result and the probability p, that two users have the same MHC profile for different number of test fields on the lateral flow assay N = {1..10} in accordance with one embodiment of the present invention.

Figure 6 depicts in a general manner three different embodiments of the test kit of the invention. The first embodiment is depicted at the top of Fig. 6 and corresponds to a test kit in the "sandwich format". Two different embodiments of the "competitive format" are depicted in the second and third row of Fig. 6, respectively. The sample (e.g. saliva or urine) containing an analyte is applied to the proximal sample application zone of a test strip. The sample containing the analyte then migrates into the distal direction through the conjugation zone or release zone, respectively, where either anti-analyte antibody or labelled analyte may be deposited. At the distal detection zone capture antibodies or analyte may be immobilized. In the further distal control zone control agents are immobilized capable of binding to the labelled antibody or labelled analyte, respectively. Excess liquid may be absorbed by a most distal absorbent pad. The embodiments depicted in Fig. 6 are in no way limiting but only illustrate certain forms of the subject matter claimed. The various zones and agents shown are not drawn to scale.

DETAILED DESCRIPTION OF THE INVENTION

The test kit according to the present invention allows for a user-friendly rapid determination of an individual's MHC profile. The term "rapid" as used herein refers to the time span between the application of a test sample and the availability of the assay result, which may range from about 1 second to about 30 minutes, preferably from about 1 minute to about 20 minutes, more preferably from about 2 minutes to about 15 minutes, most preferably from about 3 to about 8 minutes. The rapid assay can in principle be any device, kit or test format that allows the detection of MHC components within the above mentioned time span. For instance, the rapid assay may be a device based on lateral flow immunochemical technique, semiconductor technique, or any other technical device able to rapidly detect MHC compounds or a correlate thereof.

The term "user-friendly" as used herein refers to the non-requirement of any specialized skills or technical training in order to use the the test kit of the invention. The test kit of the invention is suitable for use in home and is intended to give a result which is rapid and which requires the minimum degree of skill and involvement from the user. The test kit preferably requires that some portion of the kit is contacted with the sample (e.g. a urine stream or saliva) and thereafter, no further actions are required by the user before a result can be observed. Ideally, the analytical result is observable within a matter of minutes following sample application, e.g. 30 minutes or less, preferably 10 minutes or less.

The test kit of the invention is suitable for rapidly determining an individual's MHC profile. The term "MHC profile" as used herein refers to a pattern consisting of the presence or absence of a plurality of different MHC antigens expressed by an individual.

The terms "MHC antigen" (Major Histocompatibility Antigen) and "HLA antigen" (human leukocyte-associated antigen) are used interchangeably herein. Unless specified otherwise herein, the term "MHC antigen" refers to serologically defined HLA antigens. MHC antigens in the sense of this invention include, but are not limited to, HLA-A, -B, -C, -DR and -DQ antigens as defined in [40] in Tables 1 to 6 as "WHO assigned type". The term "MHC antigens" further includes HLA antigens corresponding to the alleles listed in the Scientific Database at http://www.anthonynolan.org.uk/HIG/lists/class1list.html or http://www.anthonynolan.org.uk/HIG/lists/class2list.html. Suitable MHC antigens may also be found in [45] or at the URL http://www.allelefrequencies.net. The designations of MHC antigens with and without the prefix "HLA-" are used interchangeably herein. For example, the term "DRB1*0403" is to be understood as meaning "HLA- DRB1^*0403" and the like.

The MHC profile determined in accordance with this invention may contain information on N different MHC antigens, wherein N is an integer greater than 3. The inventors have found that it is possible to obtain sufficient information on an individual's MHC profile by analyzing a limited number of MHC antigens. Accordingly, N is preferably an integer from 4 to 40, more preferably from 5 to 33, more preferably from 6 to 30, more preferably from 6 to 25, even more preferably from 7 to 20, most preferably from 8 to 15, or from 8 to 12 (e.g. 8, 9, 10, 11 or 12).

In particular, the inventors applied a unique frequency-based selection technique for selecting suitable MHC antigens to be included in the MHC profile. In one embodiment, the set of MHC antigens to be determined includes N different MHC antigens the average phenotype frequency of which is from about 5% to about 95%, preferably from about 10% to about 90%. More preferably, the average frequency of the N different MHC antigens is from about 15 % to about 85 %, more preferably from about 20 % to about 80 %, more preferably from about 25 % to about 75 %, most preferably from about 30 % to about 70 %. These percentages refer to the average phenotypic frequencies of the respective MHC antigens in all investigated populations, as of the priority date of this application. These groups of MHC antigens are listed in the following.

Based on data from [45], the following MHC antigens were found to be present at an average frequency of from 5% to 95%, as of the priority date of this application:

A*01 ; A*02; A*0201 ; A^*03; A*11; A*1101; A*24; A*2402; B^*07; B^*0702; B^*08; B^*0801; B^*13;

B^*1301 ; B*14; B*1402; B*15; B*1501; B*1525; B*18; B^*1801 ; B^*27; B^*35; B^*3501 ; B^*38;

B^*39; B^*3901 ; B^*40; B^*4001 ; B^*4002; B*44; B^*4402; B^*4403; B^*48; BM801 ; B^*49; B^*51 ;

B*5101 ; B*52; B^*55; B^*5502; B*57; B*58; B*5801; Cw^*01 ; Cw*0102; Cw^*02; Cw^*03; Cw^*0303; Cw^*0304; Cw^*030401 ; Cw*04; Cw^*0401 ; Cw^*0403; Cw^*05; Cw^*0501 ; Cw^*06;

Cw^*0602; Cw^*07; Cw^*0701; Cw*0702; Cw^*08; CW0801; Cw^*12; Cw^*1202; Cw^*1203;

Cw*15; Cw*16; Cw*1601; DPB1*0101 ; DPB1*0201; DPB1^*0301 ; DPB1*0401 ; DPB1^*0402;

DPB1*0501 ; DPB1^*1301 ; DPB1^*1401; DQA1*0101 ; DQA1^*0102; DQA1*0103; DQA1^*0104;

DQA1*0201 ; DQA1*03; DQA1*0301 ; DQA1*0401 ; DQA1*05; DQA1*0501 ; DQB1^*02; DQB1*0201; DQB1*0202; DQB1*0301; DQB1*0302; DQB1*0303; DQB1^*0402;

DQB1*0501 ; DQB1*0502; DQB1*0503; DQB1*06; DQB1^*0601; DQB1^*0602; DQB1^*0603;

DQB1*0604; DRB1*01 ; DRB1^*0101 ; DRB1*0102; DRB1^*03; DRB1*0301 ; DRB1^*04;

DRB1*0401 ; DRB1*0403; DRB1*0404; DRB1*0405; DRB1*0406; DRB1*07; DRB1^*0701 ;

DRB1*08; DRB1*0802; DRB1*080302; DRB1*09; DRB1*090102; DRB1*11 ; DRB1^*1101 ; DRB1^*110101; DRB1^*1104; DRB1*12; DRB1M201; DRB1^*120201 ; DRB1^*13; DRB1^*1301;

DRB1*1302; DRB1^*14; DRB1^*1401 ; DRB1*1405; DRB1^*15; DRB1^*1501 ; DRB1^*150101 ;

DRB1*1502; DRB1M6; DRB1*1601 ; and DRB1*160201. This group of MHC antigens is referred to hereinafter as "5/95 group of MHC antigens".

Based on data from [45], the following MHC antigens were found to be present at an average frequency of from 10% to 90%, as of the priority date of this application:

A*01 ; A*02; A*0201; A*03; A*11; A*1101 ; A*24; A^*2402; B^*07; B^*0702; B*08; B^*0801; B^*13;

B*1301; B*14; B*15; B*1525; B*18; BM801; B^*35; B^*3501; B^*39; B*3901; B^*40; B^*4001; BM002; B*44; B*4402; B*4403; B*48; B*4801 ; B*51; B^*5101 ; B*55; B*5502; B*58; B^*5801 ; Cw*01; Cw*0102; Cw^*03; Cw^*0303; Cw*0304; Cw*030401; Cw^*04; Cw^*0401; Cw^*0403; Cw*05; Cw*0501; Cw*06; Cw^*0602; Cw*07; Cw*0701; Cw*0702; Cw*08; Cw^*0801; Cw^*12; Cw^*1202; CwM203; CwM6; DPB1*0101; DPB1^*0201; DPB1^*0301; DPB1^*0401; DPB1^*0402; DPB1^*0501; DPB1M301; DQA1*0101; DQA1^*0102; DQA1^*0103; DQA1^*0104; DQA1^*0201; DQA1^*03; DQA1^*0301; DQA1*0401; DQA1^*05; DQA1^*0501; DQB1^*02; DQB1*0201; DQB1*0202; DQB1*0301; DQB1*0302; DQB1*0303; DQB1^*0402; DQB1^*0501; DQB1^*0502; DQB1*06; DQB1*0601; DQB1^*0602; DQB1^*0603; DQB1^*0604; DRB1*01; DRB1*0101; DRB1*03; DRB1*0301; DRB1*04; DRB1*0401; DRB1^*0403; DRB1*0404; DRB1*0405; DRB1*07; DRB1*0701; DRB1*08; DRB1*0802; DRB1^*080302; DRB1*09; DRB1*090102; DRB1*11; DRB1*1101; DRB1*110101; DRB1^*1104; DRB1^*12; DRB1M20201; DRB1^*13; DRB1*1301; DRB1*1302; DRB1*14; DRB1^*1401; DRB1^*15; DRB1^*1501; DRB1^*150101; DRB1*1502; DRB1*16; DRB1^*1601; and DRB1^*160201. This group of MHC antigens is referred to hereinafter as "10/90 group of MHC antigens".

Based on data from [45], the following MHC antigens were found to be present at an average frequency of from 15% to 85% as of the priority date of this application:

A^*01; A^*02; A^*0201; A^*03; A^*11; AM 101; A^*24; A^*2402; B^*07; B^*0702; B^*08; B^*0801; BM 3;

BM 301; BM 5; BM 525; B*35; B*39; B*3901; BMO; B*4001; B*4002; B*44; B*48; BM801; Cw^*01; Cw^*0102; Cw^*03; Cw^*0304; Cw*030401; Cw*04; Cw^*0401; Cw^*0403; Cw^*0501;

Cw^*06; Cw^*0602; Cw^*07; Cw^*0701; Cw*0702; Cw*08; Cw*0801; DPB1^*0201; DPB1^*0301;

DPB1^*0401; DPB1^*0402; DPB1^*0501; DQA1*0101; DQA1*0102; DQA1^*0103; DQA1^*0201;

DQA1^*03; DQA1^*0301; DQA1^*0401; DQA1^*05; DQA1^*0501; DQB1^*02; DQB1^*0201;

DQB1^*0202; DQB1^*0301; DQB1*0302; DQB1*0402; DQB1^*0501; DQB1^*06; DQB1^*0601; DQB1^*0602; DRB1^*01; DRB1*03; DRB1^*0301; DRB1^*04; DRB1^*07; DRB1^*0701;

DRB1^*08; DRB1*0802; DRB1*080302; DRB1*09; DRB1*090102; DRB1M1; DRB1M10101;

DRB1M2; DRB1M20201; DRB1M3; DRB1M4; DRB1M401; DRB1M5; DRB1M501;

DRB1M50101; and DRB1M60201. This group of MHC antigens is referred to hereinafter as

"15/85 group of MHC antigens".

Based on data from [45], the following MHC antigens were found to be present at an average frequency of from 20% to 80%, as of the priority date of this application:

A*01; A*02; A*0201; A*03; AM1; AM 101; A^*24; A*2402; BM301; B^*35; B^*3901; B^*40; B*4001; B*44; Cw*03; Cw*030401; Cw*04; Cw*0401; Cw^*07; Cw^*0701; Cw^*0702; Cw*0801 ; DPB1*0201 ; DPB1*0401 ; DPB1*0402; DPB1*0501 ; DQA1^*0102; DQA1^*03; DQA1^*0301; DQA1^*05; DQA1^*0501 ; DQB1*02; DQB1^*0201 ; DQB1^*0301 ; DQB1^*0302; DQB1^*06; DRB1^*0301 ; DRB1*04; DRB1*07; DRB1*0701 ; DRB1^*080302; DRB1^*11 ; DRB1^*110101 ; DRB1M2; DRB1^*120201; DRB1*13; DRB1*14; and DRB1*15. This group of MHC antigens is referred to hereinafter as "20/80 group of MHC antigens".

Based on data from [45], the following MHC antigens were found to be present at an average frequency of from 25% to 75%, as of the priority date of this application:

A^*01 ; A^*02; A^*0201; A^*11 ; A^*1101; A*24; A*2402; B*1301 ; B^*3901 ; B^*4001 ; Cw^*03; Cw*030401 ; Cw*07; Cw*0701 ; Cw*0702; DPB1*0201 ; DPB1^*0401 ; DPB1^*0402; DQA1^*0102; DQA1^*03; DQA1^*0301 ; DQA1*05; DQA1^*0501 ; DQB1^*02; DQB1^*0201; DQB1^*0301 ; DQB1^*06; DRB1^*04; DRB1^*080302; and DRBT120201. This group of MHC antigens is referred to hereinafter as "25/75 group of MHC antigens".

Based on data from [45], the following MHC antigens were found to be present at an average frequency of from 30% to 70%, as of the priority date of this application:

A^*02; A^*0201 ; A^*11 ; A^*1101 ; A*24; A*2402; Cw*03; Cw*030401 ; Cw^*07; DPB1^*0201 ; DPB1*0401 ; DQA1*0102; DQA1*03; DQA1*05; DQA1*0501 ; DQB1*02; DQB1^*0301 ; and DQB1*06. This group of MHC antigens is referred to hereinafter as "30/70 group of MHC antigens".

In one embodiment, all MHC antigens to be determined with the test kit of the invention are selected from one of the above groups of MHC antigens. By selecting MHC antigens according to the frequency in the population, a significant result can be obtained without having to analyse a vast number of possible MHC alleles on the level of the genome. The frequency of an MHC antigen is defined as the proportion with which a given antigen can be detected in a given population. A population can, for example, be a geographically defined group of individuals, for example human individuals. As used herein, the frequency of an MHC antigen is identical to the average phenotype frequency determined on the basis of data from [45] and/or http://www.allelefrequencies.net, as of the priority date of this application. Suitable antigens include, but are not limited to, those listed in Tables 1 and 2. In a specific embodiment, the MHC antigens to be analyzed by the test kit of the invention are selected from the group consisting of HLA-A^*01 , HLA-A^*02, HLA-A^*03, HLA-AM 1 , HLA-A^*24, HLA- A^*26, HLA-A^*30, HLA-A^*31 , HLA-A^*68, HLA-B^*07, HLA-B^*08, HLA-B^*15, HLA-B^*35, HLA- BMO, HLA-B*44, HLA-B*51 , HLA-Cw*01 , HLA-Cw^*03, HLA-Cw^*04, HLA-Cw^*07, HLA- DPB1^*0201 , HLA-DPB1^*0301 , HLA-DPB1*0401 , HLA-DQA1^*0101 , HLA-DQA1^*0102, HLA- DQA1^*0201 , HLA-DQA1*03, HLA-DQA1*0301 , HLA-DQA1*05, HLA-DQA1^*0501 , HLA- DQA1^*0505, HLA-DRB1*01 , HLA-DRB1*03, HLA-DRB1^*0301, HLA-DRB1^*04, HLA- DRB1*07, HLA-DRB 1^*0701 , HLA-DRB1*11, HLA-DRB1*13, HLA-DRB1*15, and HLA- DRB1^*1501. In another embodiment, the set of MHC antigens to be analyzed comprises both class I and class Il MHC antigens.

In order to determine whether or not an individual has a certain MHC antigen (i.e. whether or not he/she expresses it), the presence or absence of an analyte in a test sample from said individual is detected in accordance with this invention.

The term "test sample" refers to a material suspected of containing the analyte. The test sample may, for instance, include materials derived from a biological source, such as a physiological fluid, including, saliva, sweat, urine, blood, interstitial fluid, plasma, ocular lens fluid, cerebral spinal fluid, milk, ascites fluid, mucous, synovial fluid, peritoneal fluid, vaginal fluid, menses, amniotic fluid or the like. The body fluid may be diluted or processed prior to applying it in the test kit.

As used herein, the term "analyte" refers to the substance to be detected. The analyte which is suspected of being present in the test sample and which is to be detected may be any substance indicative of a specific MHC antigen.

MHC molecules, their fragments, degradation products of their peptide ligands and products of MHC-dependent microflora have all been considered as potential odorants. The most significant contacts between peptides and the MHC molecules are mediated through the side chains of so-called anchor residues of the peptide fitting into pockets of the MHC molecules. The range of peptides displayed by the MHC molecules of an individual mirrors the structural diversity of its MHC alleles. Different peptide ligands of one particular MHC molecule share common residues (so-called anchors) whose side chains fit into the characteristic binding pockets of the MHC molecule [25]. This suggests that anchor residues could be the defining feature of peptides and olfactory assessment might focus on them. MHC-peptide complexes are shed from the cell surface and their fragments appear in serum, saliva, sweat and urine [41]. The molecular weight for MHC membrane proteins and peptide ligands found in human saliva is in the range of 40 kD [12] and similar proteins have been detected in urine and sweat. Soluble HLA molecules between 45 kD (class I heavy chain) and 23 kD have been found in urine and 45 kD and 40 kD soluble HLA associated molecules in sweat.

Preferably, the analyte is selected from the group consisting of MHC molecules, fragments of MHC molecules, MHC ligands, fragments of MHC ligands, and combinations thereof. For example, the analyte may be a peptide ligand which can be bound by its respective MHC molecule in the molecular groove. Alternatively, the peptide may already have been partially degraded resulting in fragments which may be detected as well. In body fluids such as saliva and urine, full length MHC molecules may be present. Usually, however, fragments of MHC molecules such as the extracellular portion thereof or fragments thereof are present in the test sample. It is also possible to detect a combination of MHC molecule and its ligand bound thereto. For instance, analytes may be MHC (major histocompatibility) components. These compounds may be part of the MHC gene complex or correlated to it and may include MHC-molecules, MHC-ligands, MHC-correlated peptides, MHC-correlated odour components, MHC-derived odour components, MHC-correlated carboxylic acids or antibodies to any of the above substances. In a particular embodiment, the analyte is selected from the group consisting of MHC molecules, fragments of MHC molecules, naturally occurring derivatives of MHC molecules, MHC ligands, fragments of MHC ligands, naturally occurring derivatives of MHC ligands and combinations thereof. The analyte may be a peptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1 to 618.

The test kit may be any kit or device suitable for rapidly determining an MHC profile of an individual. The analyte can be detected in a "sandwich" or in a competition assay. The general principle of such test formats is known to the skilled person. Particularly preferred are lateral flow assays as described in e.g. EP 291 194 B1 , EP 186 799 B1 , or WO 2005/121800 A2. The general principles of the test kits, test devices and assay methods disclosed in US 4,956,302; US 6,372,513; US 5,770,460 and/or US 4,943,522 can be applied in the present invention. Sandwich format

The so-called sandwich format of the test kit is preferred when the analyte is an MHC receptor, a soluble form thereof or a fragment thereof.

This assay format typically involves a porous media having a mobilizable labeled antibody and an immobilized unlabeled antibody partner for the analyte of interest in the biological sample. These antibodies are often referred to as the conjugate antibody and the capture antibody. The sample is added to the porous media, to allow for formation of labeled mobilizable product which moves along the porous media to contact and react with the capture antibody to form a fixed, detectable, concentrated capture antibody/analyte/detection reagent complex. Sandwich assays may include immunological assays wherein the labeled reagent and the second binding partner are both antibodies, or are both antigens, and may also include other types of molecules. For example, enzyme immunoassays (EIA) and enzyme-linked immunosorbent assays (ELISA) are types of sandwich immunoassays, in which the binding is between an antibody and an antigen, and the labeling partner is an enzyme.

Typically, these chromatographic assay devices are comprised of a porous chromatographic medium which acts as the matrix for the binding assay. The sample of interest is added directly or indirectly to one end of the medium, and is chromatographically transported to a detection reagent with which it reacts to form a labeled conjugate product, which is then transported to a test zone containing an immobilized capture reagent such as a capture antibody, in which the presence, absence, or quantity of an analyte of interest can be determined.

The preferred test kit of the invention comprises a plurality of different pairs of binding reagents; usually it comprises N mobilizable labelled primary binding reagents and N immobilizable unlabelled secondary binding reagents, wherein N is an integer as defined hereinabove. The binding reagent may be any molecule capable of binding to the analyte. Preferably, the binding reagent is an antibody, e.g. a monoclonal antibody or a polyclonal antibody. As used herein, the term "antibody" is meant to include intact molecules as well as antibody fragments (such as, for example, Fab and F(ab')₂ fragments) which are capable of specifically binding to the analyte. Fab and F(ab')₂ fragments lack the F_c fragment of intact antibody. Moreover, antibodies in accordance with the present invention include single chain antibodies. Other binding reagents such as aptamers may also be envisaged as long as they are capable of specifically binding to the analyte.

MHC class I antigen-antibody conjugates have been produced for rats (A^c with YR5/12 and A^av1 with JY1/116) [41] and humans (class I associated proteins with 4B5.1 monoclonal primary antibody and IgG phosphatase-conjugated goat anti-mouse or anti-rabbit secondary antibody) [29]. Suitable antibodies specifically recognizing the analyte can be generated or otherwise obtained by the skilled person. Suitable techniques for generating antibodies are disclosed in, e.g., Harlow and Lane, "Antibodies, A Laboratory Manual" CSH Press 1988, Cold Spring Harbor N.Y. Specifically, the following examples for commercially available antibodies for the sandwich type assay shall visualize the manufacturing of such a test kit (See Figure 6): Control antibodies can be labeled rat anti mouse IgGI heavy chain:HRP (AbD Serotec No. MCA336P) or mouse anti human beta 2 microglobulin (AbD Serotec No. MCA 1116) labeled with Lynx rapid HRP antibody conjugation Kit (AbD Serotec No. LNK002P). Detection (capture) antibodies can for example be mouse anti human HLA A2 (AbD Serotec No. MCA2090), mouse anti human HLA A24 (antikoerper-online.de, No. ABIN131788), mouse anti human HLA B7 (AbD Serotec No. MCA986), mouse anti human HLA DQw1 (antikoerper-online.de, No. ABIN100554) mouse anti human HLA DRA (antikoerper-online.de, No. ABIN228532), mouse anti human HLA DR (AbD Serotec No. MCA1879) etc.

Thus, for each MHC-antigen indicating analyte, two binding reagents specifically recognizing the analyte are included. This allows a more sensitive detection of the analyte. A given pair of binding reagents consists of (i) an unlabelled secondary binding reagent that is capable of specifically binding to an analyte that is indicative of a predefined MHC antigen, and (ii) a labelled primary binding reagent that is capable of specifically binding to the same analyte. In the case of antibodies as binding reagents, the two antibodies of a pair of antibodies preferably recognize different epitopes present in the analyte molecule(s). Each pair of binding reagents contained in the kit usually detects an MHC antigen different from the MHC antigens detected by the other pairs of binding reagents in the same kit. Accordingly, a test kit containing N pairs of binding reagents will allow detection of N different MHC antigens, respectively.

With reference to the frequency-based selection of MHC antigens to be analysed by the test kit of this invention, the test kit preferably comprises N different pairs of binding reagents capable of specifically recognizing N different MHC antigens, respectively, wherein said N different MHC antigens are selected from the 10/90 group of MHC antigens.

More preferably, the test kit comprises N different pairs of binding reagents capable of specifically recognizing N different MHC antigens, respectively, wherein said N different MHC antigens are selected from the 15/85 group of MHC antigens.

Still more preferably, the test kit comprises N different pairs of binding reagents capable of specifically recognizing N different MHC antigens, respectively, wherein said N different MHC antigens are selected from the 20/80 group of MHC antigens.

Even more preferably, the test kit comprises N different pairs of binding reagents capable of specifically recognizing N different MHC antigens, respectively, wherein said N different MHC antigens are selected from the 25/75 group of MHC antigens or from the 30/70 group of antigens.

Competitive format

The so-called competitive format of the test kit is preferred when the analyte is an MHC ligand, a fragment or a derivative thereof.

In a first embodiment, the test kit in the competitive format comprises: at least one solid substrate in contact with a proximal sample application zone, said substrate having N conjugation zones and N distal detection zones, said conjugation zones respectively containing N labelled primary binding reagents capable of binding with an analyte to form an analyte-primary binding reagent conjugate; said distal detection zone respectively having immobilized thereunto, N unlabeled secondary binding reagents capable of binding to the respective labelled primary binding reagent, but not to a migrating analyte-labeled primary binding reagent conjugate to determine the presence or absence of N analytes which are indicative of defined N-MHC antigens, wherein N is an integer greater than 3.

In this first embodiment, the labelled primary binding reagent is preferably a labelled antibody specifically recognizing the analyte. The secondary binding reagent is preferably an immobilized analyte which is in the detection zone. The labelled antibody can bind to the immobilized analyte, while the labelled antibody cannot bind to the immobilized analyte when it is bound to the analyte in an analyte-antibody complex. Thus, the migrating analyte competes with the immobilized analyte for binding to the labelled antibody. The lack of a signal in the detection zone is indicative of the presence of a given analyte in the sample.

In a second embodiment, the test kit in the competitive format comprises: at least one solid substrate in contact with a proximal sample application zone, said substrate having N release zones and N distal detection zones, said release zones respectively containing N different labelled analytes; said distal detection zone respectively having immobilized thereunto, N unlabeled binding reagents capable of binding to a migrating labelled or unlabelled analyte to determine the presence or absence of N analytes which are indicative of defined N-MHC antigens, wherein N is an integer greater than 3.

In this second embodiment, the unlabelled binding reagent immobilized in the detection zone is preferably an immobilized antibody capable of specifically recognizing the analyte.

The release zone preferably contains labelled analyte molecules which can migrate when sample liquid migrates from the sample application zone through the release zone into the direction of the detection zone. The unlabelled analyte from the sample then competes with the labelled analyte from the release zone for binding to the immobilized unlabelled binding reagents. The lack of a signal in the detection zone is indicative of the presence of a given analyte in the sample.

Tables 1 and 2 respectively show selection of MHC class I and class Il alleles with corresponding peptide ligands to obtain the desired result for individual MHC profiles from lateral flow assay. Bold letters within selected ligands mark anchor residues likely to bind to groves of the corresponding MHC membrane protein. Tables 3-22 show further MHC ligands which may be detected as analytes in the sample, being indicative of the respective MHC antigens.

Test kit design

The test kit of the instant comprises one or more solid substrates. A preferred substrate of the present invention is a porous medium, strip or membrane. In one embodiment, the test kit comprises one solid substrate which in turn comprises said at least 3 different pairs of binding molecules. In another embodiment, the test kit comprises 2 or more solid substrates, e.g. at least 3 solid substrates, 8 to 33 solid substrates or 8 to 12 solid substrates. In a specific embodiment, the number of solid substrates in the test kit is identical to the number of pairs of binding reagents comprised in the test kit, i.e. the test kit comprises N solid substrates. Suitable solid substrate materials include, but are not limited to, cellulose, nitrocellulose, polyethylene, polyvinyl chloride, polyvinyl acetate, copolymers of vinyl acetate and vinyl chloride, polyamide, polycarbonate, polystyrene; and "bibulous" materials such as those disclosed in US Patent No. 5,770,460 treated with blocking agents, e.g. with detergents or proteins. Suitable solid substrate materials are known to one of ordinary skill in the field.

Referring to Figure 2a, the solid substrate may be a lateral flow membrane, which has on one end, a proximal sample application zone to receive a liquid sample, and further upstream, a conjugation zone wherein the analyte conjugates with a mobilizable labeled primary binding reagent, and further upstream, at least one distal detection zone having an immobilized unlabelled binding reagent affixed thereunto. The test sample contains an analyte which is an analyte which can be derivatized so as to bind the immobilized member. In one preferred configuration, the membrane is bound to two substantially fluid-impervious sheets, one on either side, with openings on one side or both sides to provide definition to the application and detection zones.

The lateral flow achieved in the kit and method of the invention is the result of the properties of the lateral flow membrane. The membrane has a much smaller thickness than surface dimension and may be hydrophilic enough to be wetted and thus permit aqueous solutions and materials to exhibit lateral flow freely, and preferably isotropically, at substantially the same rates for various components of a sample.

The membrane usually includes more than one detection zone (see Figure 2a and Example 1), usually it includes N detection zones, along with control and reference zones. Multiple detection zones are designed to detect different analytes indicative of different MHC antigens. Multiple detection zones may be in any spatial relationship to the application zone, since the membrane itself does usually not provide a barrier to sample flow.

In the method of the invention, the sample, to be analyzed for MHC-antigen indicating analytes is applied to the proximal sample application zone and allowed to be transported laterally through the membrane via a conjugation zone containing the first member of the binding reagent pair to a distal detection zone, where there is, immobilized on the membrane, the other, second member of the binding reagent pair.

In the detection zone, mobilizable binding reagent-analyte conjugate binds to the immobilized binding reagent, and the resulting bound complex is detected. Detection may use any of a variety of labels and/or markers, e.g., enzymes, liposomes, fluorescent tags, polymer dyes, or colored particles, etc., and detection is by means of, for example, direct visual observation, by developing a color, by fluorescence measurement, or by any of many other techniques by which the presence or absence of a chemical or biochemical species may be detected directly or indirectly.

The kit may be encased in a housing. The housing can be fabricated out of any convenient material (e.g. HDPE, LDPE, PP, polystyrene, acrylic, polycarbonate, etc.).

The rapid assay to which a test sample is applied is designed such that it can measure a set of different analytes that are present within one test sample. The number of different analytes may be N as defined hereinabove, preferably it is 8 - 12. Each detected analyte contributes to the assay result.

It is preferred that the test kit of the invention comprises an item or article on which a unique identifier is recorded. The unique identifier may be any information suitable to uniquely characterize the test kit. The unique identifier may be a number such as a serial number consisting of several digits (for example 5, 6, 7, 8, 9, 10, 11 or 12 digits), a barcode or a graphic representation which can be identified by a scanner.

The item on which the unique identifier is recorded may be any article, product or the like suitable to carry such information. It is preferred that the unique identifier is imprinted on said item or article. Preferably, the item or article is made of paper or cardboard. In another preferred embodiment, the unique identifier is imprinted on the solid substrate of the test kit. In yet another preferred embodiment, the unique identifier is imprinted on a housing in which the solid substrate is encased. To obtain an encrypted MHC profile, the configuration of the sensors, the signals or the substrates in the rapid assay may be encrypted. The key, which is related to said unique identifier, together with the encrypted MHC profile yields a decrypted MHC profile. The unique identifier may be a serial number or a random number without replacement or any other unambiguous number. This number may be indicated on the rapid assay. One embodiment of the present invention comprises the steps of obtaining a plurality of personal profiles from a plurality of individuals and storing said personal profiles in a database on a computer. The personal profile may encompass contact data such as postal address, telephone number, e-mail address of the individual. The personal profile may further include a photograph or image of the individual as well as personal attributes and preferences (e.g. height, colour, gender, weight, activities and the like). The personal profiles may be entered by the individual into a form on a computer screen provided by a first service provider, e.g. an internet dating service. There may be several fields which must be filled and further fields which may optionally be filled. By confirming the entry of his/her personal profile by clicking on a virtual button on the computer screen, the personal profile is transmitted to a first service provider which may be the dating service. Once the personal profile is received by the service provider, it is stored in a database on a computer. Known forms of suitable databases are known of ordinary skill in the field.

Alternatively, the personal profile may be transmitted by e-mail, by normal mail, via telefax or over the telephone.

In a further step, a test kit for rapidly determining an individual's MHC profile is provided to those individuals who have already provided their personal profiles. Alternatively, the test kit may be offered to interested individuals who have not yet provided their personal profiles. If interested, they may use the kit, determine their encrypted MHC profile and enter the results together with their personal profiles into the platform of the dating service, or enter the encrypted MHC profile in the personal data separately.

Preferably, the test kit is a test kit according to the present invention as described herein above.

Referring to Figure 1 , one embodiment of the invention can be summarized as follows: A user from a defined community 2 receives from a dating service a rapid assay 3 to detect an encrypted MHC profile. Said user performs the rapid assay and adds the results of the rapid assay (the encrypted MHC profile) on an interface of the dating service 4 to the personal profile of said user together with a unique identifier indicated on the rapid assay. The dating service 5 sends the results and the unique identifier 6 to a matching service 8. The matching service converts the assay results using a key, which is related to said unique identifier, into a decrypted MHC profile. The pair wise distance between decrypted MHC profiles is then converted into a quality of match, where intermediate distances between decrypted MHC profiles constitute better matches than large or small distances. The quality of match is sent to the dating service 7 from where said user can acquire the quality of match between said user and any other user with an account at the dating service and a valid entry for the assay result and unique identifier. It should be understood, that the company shipping the rapid assay to the user is preferably different from the matching service but can be identical to the dating service or to any other service. The company shipping the rapid assay to the user may even be the manufacturer.

Together with the encrypted MHC profile, the respective unique identifier is transmitted from the user to the first service provider. Accordingly, the entry form on the computer screen provided by the dating service is preferably adapted to allow the entry of two pieces of information, namely (1) the encrypted MHC profile and (2) the unique identifier. Preferably, the filling of both fields is mandatory. Entry of only one of both pieces of information would not allow transmission of the data.

The encrypted MHC profiles and the respective unique identifiers are stored in a database, preferably in a database on a computer. Each encrypted MHC profile and its corresponding unique identifier is allocated to one of the personal profiles.

The encrypted MHC profiles are decrypted using the decryption key, which is related to said unique identifier, preferably by the aid of a computer. The thus decrypted MHC profiles are than stored in a database, preferably in a database on a computer.

The matching of the decrypted MHC profiles involves two steps. Namely the step of calculating a distance measure and the transformation of said distance measure into a quality of match. The distance measure is a metric or distance function which defines a distance between elements of a set. Here we refer to the mathematical definition. Specifically a metric on a set X is a function d : X * X → R (where R is the set of real numbers). For all x, y, z in X, this function is required to satisfy the following conditions: 1. Cl(X₁ y) >0

2. d(x, y) = 0 if and only if x = y

3. d(x, y) = d(y, x) 4. d(x, z) <d(x, y) + d(y, z)

The distance measure may for instance be a Euclidian distance, a Mahalanoby distance or any other metric suitable for the MHC resolution addressed by the detected analytes. The distance measure may include the phylogeny of the MHC genes coding for the detected analytes such as the measures for example developed by or based on the following authors [42-44].

The next step thus comprises calculating the pairwise distances in decrypted MHC profiles between all individuals for which decrypted MHC profiles have been stored in the database. Theses results can be used to generate a matrix that contains the quality of match for each pairwise distance as depicted in Figure 4. From the pairwise distances, the quality of match between individuals can be calculated, preferably using a computer.

The quality of match may be defined as being "high" or "low". The quality of match between two individuals can be defined as "high" if more than 10 % and less than 90 % of the MHC antigens in the MHC profiles of the two individuals are the same. Preferably, the quality of match is regarded as "high" if more than 15 % and less than 85 % of the MHC antigens in the MHC profiles of the two individuals are the same. More preferably, the quality of match is regarded as "high" if more than 25 % and less than 75 % of the MHC antigens in the MHC profiles of the two individuals are the same.

The method of the invention may further comprise the step of providing to a first individual contact data of further individuals whose MHC profiles have a high quality of match with the MHC profile of said first individual. These data are usually provided only upon request by the first individual. The request may be submitted "online" by clicking on a virtual button or link on the computer screen.

In a specific embodiment, steps (a), (b), (c) and (d) are carried out by a first service provider while steps (e), (f) and (g) are carried out by a second service provider. That is, the decryption of the MHC profile is carried out by a different entity than that having contact with the user. For example, steps (e), (f) and (g) may be carried out by a so called matching service whereas steps (a) through (d) are carried out by the dating service which offers to provide a match between individuals.

The test kit of this invention may also be used for determining the degree of match between an individual wishing to adopt a child, and a child. The preferred embodiments of the test kit described herein apply to this use mutatis mutandis.

The following examples further illustrate the invention. The invention, however, is not limited to these exemplary embodiments.

EXAMPLES

Example 1: Predicting the matching quality between users based on an MHC flow- through assay

In a first step, the user enters their personal profile into an online form. The personal profile comprises among other things the postal address of said user. Referring to Figure 2b, for instance, it may also comprise a personal photograph 24. The personal profile of said user is then sent to the dating service. The dating service stores the personal profile into its database and sends a rapid assay kit to the postal address of said user.

A user performs the rapid assay. Referring to Figure 2a, for instance, one embodiment of a rapid assay in our invention may be a flow-through assay device 9. To initiate the flow through assay, a user directly applies approximately 5 ml of saliva sample to a sample pad

10 through which it will travel in the direction of the wicking pad 15. In the conjugate zone

11 , a gold colloid labelled primary antibody will bind to the analyte. The superfluous conjugate will travel further through the porous membrane made of nitrocellulose 12 to the detection region 13, where a secondary antibody binds to the conjugate. The primary and the secondary antibodies are designed and manufactured such that they detect MHC antigens with two epitopes. The sensitivity of the antibodies to detect MHC antigens in the saliva sample is approximately 50 ng ml^'1. Superfluous conjugate moves further to the control region 14 where an anti-species antibody binds to the conjugate. The rest of the conjugate will be absorbed by the wicking pad 15, which promotes capillary action and fluid flow through the porous membrane 12. Staining in the detection region 13 and the control region 14 together indicate the presence or absence of each of the nine analytes for which the flow through device has been designed. A serial number 18 is indicated on the rigid material 16 containing the device 9. Presence and absence of each of the nine analytes together constitute an encrypted MHC profile of the user.

Referring to Figure 2b, the user enters the encrypted (ER) MHC profile 20 and the serial number 21 into the online form and sends it to the dating service 23. The dating service stores the encrypted MHC profile and the serial number of said user and sends it to the matching service.

The matching service receives the encrypted MHC profile and the serial number of said user. In order to decrypt the encrypted MHC profile of said user, the key to the corresponding serial number is sorted out from the key table in Figure 4 and applied to the encrypted MHC profile of said user, which yields the decrypted (DR) MHC profile of said user. Then the decrypted MHC profile of said user is stored in the MHC profiles table (see Figure 4). In the next step, the distance-vector of said user to the decrypted MHC profiles of all other users is computed and the distance matrix in Figure 4 is updated with this distance vector.

The distance vector of said user is then transformed into a match-quality vector of said user. To compute the match-quality vector of said user, a function is applied to transform the distances of the distance-vector to match-quality parameters between 0 and 1 where 0 represents the lowest and 1 the best match-quality. The matrix that contains the quality of match is then updated with the vector that contains the quality of match of said user to all other users. Then the matching service sends the match-quality vector of said user to the dating service in order to protect said user's privacy, by separating said user's personal profile from said user's decrypted MHC profile. The dating service receives the vector of said user and updates its matrix.

Referring to Figure 2b, in the next step said user enters the desired matching quality 22 and other preferences into the online form and sends the search criteria to the matching service 23. The search criteria are then processed and the necessary information is retrieved from the database of the matching service. Then the matches are computed and send to said user where they are displayed. Said user checks the matches and then either ends the search for a dating partner or performs a new search.

By way of example two possible ways how to calculate the "match-quality matrix" on the basis of the "MHC profiles" via the "distance matrix" are indicated in the following:

Triangle-Curve (see Example 1)

, ≤ N

Quadratic-Curve

r_{l k} are single (boolean) assay results which together constitute the decrypted MHC profile DR d,_j is the distance function between the decrypted MHC profiles DR, and DR_j q,_j is the function to obtain the quality of match to be entered in the match quality matrix.

Example 2: Selection of target antigens

Single MHC loci can have up to approximately 300 different alleles and most of these alleles occur in small frequencies in the population. Preferably, lateral flow assay of the instant invention has several test fields (9 in Example 1) of which each has a certain probability for a positive or negative result. If the frequency of the alleles is much higher or much lower than 0.5 in the population, then the expected diversity from individual MHC profiles will not be sufficient to effectively match the profile of two or several users with each other. To circumvent this problem, MHC class I and class Il alleles with phenotypic frequencies between 0.2 and 0.8 (which corresponds to 20% - 80%) on www.allelefrequencies.net are used [45]. Other allele combinations listed in http://www.anthonynolan.orq.uk/HIG/lists/class1list.html or http://www.anthonynolan.orq.uk/HIG/lists/class2list.html could also be a candidate. The threshold of 0.2 and 0.8 was calculated from the expected frequency necessary to distinguish individuals with a given probability

where p, is the probability with which a single test field shows a positive result, p the probability that two randomly drawn users have the same MHC profile (see Figure 5) and N the number of test fields on the assay (9 in our examples, see Figure 2a). For our examples we use 0.2 < p, < 0.8. The number of test fields is preferably greater than 5 with a preferred upper limit of 18 - 33.

It will be apparent to those skilled in the art that various modifications and variations can be made to the objects of the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents. Accordingly, the invention is not limited by the embodiments described above which are presented as examples only but can be modified in various ways within the scope of protection defined by the appended patent claims. All documents cited herein are incorporated by reference as if included in this specification in their entireties.

Table 1 : Selection of MHC class I alleles with corresponding peptide ligands to obtain the desired result for individual MHC profiles from the lateral flow assay. Bold letters within selected ligands mark anchor residues likely to bind to groves of the corresponding MHC membrane protein.

1) Data from www.allelefreauencies.net

2) Data from www.svfpeithi.de

3) Scores are based on the frequency of the respective amino acids in natural ligands, T-cell epitopes or binding peptides. For details see Rammensee etal. (1999)

Table 2: Selection of MHC class Il alleles with corresponding peptide ligands to obtain the desired result for individual MHC profiles from the lateral flow assay. Bold letters within selected ligands mark anchor residues likely to bind to groves of the corresponding MHC membrane protein.

4) Data from www.allelefrequencies.net

5) Data from www.svfpeithi.de

6) Scores are based on the frequency of the respective amino acids in natural ligands, T-cell epitopes or binding peptides. For details see Rammensee etal. (1999)

The following tables 3-22 show various ligands for a number of selected MHC antigens:

Table 3: Pe tide Li ands of HLA-A*01

OO

References

1. Wedekind, C. and D. Penn, MHC genes, body odours, and odour preferences. Nephrology Dialysis Transplantation, 2000. 15: p. 1269 - 1271.

2. Eggert, F., et al., The major histocompatibility complex and the chemosensory signalling of individuality in humans. Genetica, 1999. 104: p. 265 - 273.

3. Penn, DJ. and W. K. Potts, The evolution of mating preferences and major histocompatibility complex. The American Naturalist, 1999. 153(2): p. 145 - 164.

4. Wedekind, C, et al., MHC-dependent mate preferences in humans. Proc. R. Soc Lond. B, 1995. 260: p. 245 - 249.

5. Singh, P.B., Chemosensation and genetic individuality. Reproduction, 2001. 121: p. 529 - 539.

6. Penn, DJ. , et al., Individual and gender fingerprints in human body odour. J. R. Soc. Interface, 2006. 4: p. 331 - 340.

7. Restrepo, D., et al., Odortypes and MHC peptides: complementary chemosignals of MHC haplotype. Trends in Neurosciences, 2006. 29(11 ).

8. Milinski, M., et al., Mate choice decisions of stickleback females predictably modified by MHC peptide ligands. Prodeedings of the National Academy of Science, 2005. 102(12): p. 4414 - 4418.

9. Abies, E. M., L. M. Kay, and J. M. Mateo, Rats assess degree of relatedness from human odors. Physiology & Behavior, 2007. 90: p. 726 - 732.

10. Wyart, C, et al., Smelling a single component of male sweat alters levels of Cortisol in women. Journal of Neuroscience, 2007. 27(6): p. 1261 - 1265.

11. Milinski, M. and C. Wedekind, Evidence for MHC-correlated perfume preferences in humans. Behavioral Ecology, 2001. 12(2): p. 140 - 149.

12. Jacob, S., et al., Paternally inherited HLA alleles are associated with women's choice of male odor. Nature Genetics, 2002. 30(2): p. 175 - 179.

13. Garver-Apgar, C. E., et al., Major histocompatibility complex alleles, sexual responsivity, and unfaithfulness in romantic couples. PsychSc, 2006. 17(10): p. 830 - 836. 14. Milinski, M., The major histocompatibility complex, sexual selection, and mate choice. Annual Review in Ecology, Evolution, and Systematics, 2006. 37: p. 159 - 186.

15. Nowak, M.A., K. Tarczy-Hornoch, and J. M. Austy, Proc. Natl. Acad. Sci., 1992. 89: p. 10896.

16. DeBoer, R.J. and A.S. Perelson, Proceedings of the Royal Society London B, 1993. 252: p. 171.

17. Reusch, T.B.H., et al., Female sticklebacks count alleles in a strategy of sexual selection explaining MHC polymorphism. Nature, 2001. 414: p. 300 - 302.

18. Wegner, K.M., et al., Parasite selection for immunogenetic optimality. Science, 2003. 301: p. 1343.

19. Zerefos, P. G., et al., Characterization of the human urine proteome by preparative electrophoresis in combination with 2-DE. Proteomics, 2006. 6: p. 4346 - 4355.

20. Singer, A.G., G. K. Beauchamp, and K. Yamazaki, Volatile signals of the major histocompatibility complex in male mouse urine. Proc. Natl. Acad. Sci., 1997. 94: p. 2210 - 2214.

21. Krieger, J., et al., Selective activation of G protein subtypes in the vomeronasal organ upon stimulation wiht urine-derived compounds. The Journal of Biological Chemistry, 1999. 274(8): p. 4655 - 4662.

22. Leinders-Zufall, T., et al., MHC class I peptides as chemosensory signals in the vomeronasal organ. Science, 2004. 306(5): p. 1033 - 1037.

23. Montag, S., et al., "Electronic nose" detects major histocompatibility complex- dependent prerenal and postrenal odor components. Proc. Natl. Acad. Sci., 2001. 98: p. 9249 - 9254.

24. Boehm, T. and F. Zufall, MHC peptides and the sensory evaluation of genotype. Trends in Neurosciences, 2006. 29(2).

25. Rammensee, H.-G., et al., SYFPEITHI: database for MHC ligands and peptide motifs. Immunogenetics, 1999. 50: p. 213 - 219.

26. Yewdell, J.W. and J. R. Bennink, lmmunodominance in major histocompatibility complex class l-restircet T lymphocyte responses. Annual Review in Immunology, 1999. 17: p. 1 - 30.

27. Zavazava, N., G. Leimenstoll, and W. Mϋller-Ruchholtz, Measurement of soluble MHC class I molecules in renal graft patients: a noninvasive allograft monitor. Journal of Clinical Laboratory Analysis, 1990. 4: p. 426 - 429.

28. Adamashvili, I., et al., Soluble HLA: patterns of expression in normal subjects, autoimmune diseases, and transplant recipients. Rheumatol Int, 2005. 25: p. 491 - 500. 29. Wobst, B., et al., Molecular forms of soluble HLA in body fluids: potential determinants of body odor cues. Genetica, 1999. 104: p. 275 - 283.

30. Adamashvili, I., et al., Soluble HLA in saliva of patients with autoimmune rheumatic diseases. Rheumatol Int, 2002. 22: p. 71 - 76.

31. Hayman, M.W., et al., Soluble human leukodyte antigen: A diagnostic indicator of rheumatoid arthritis. Journal of Immunological Methods, 2006. 315: p. 19 - 26.

32. Amirghofran, Z., et al., Journal of Cancer Research Clinical Oncology. 128, 2002(443 - 448).

33. Utsumi, M., et al., Expression of major urinary protein genes in the nasal glands associated with general olfaction. Neurobiology, 1999. 39: p. 227 - 236.

34. Brennan, P.A. and F. Zufall, Pheromonal communication in vertebrates. Nature, 2006. 444: p. 308 -

35. Fan, J., et al., Visual perception of male body attractiveness. Proceedings of the Royal Society London B, 2005. 272: p. 219 - 226.

36. Tovee, M.J., et al., Effects of sexual dimorphism on facial attractiveness. Lancet, 1999. 399: p. 215 - 216.

37. Perrett, D. I., et al., Effects of sexual dimorphism on facial attractiveness. Nature, 1998. 394: p. 884 - 887.

38. Buss, D. M., The evolution of desire: strategies of human mating. 1994, New York: Basic Books.

39. Gourab Goshal, P.H., Attractiveness and activity in internet comunities. Physica, 2006. 364: p. 603 - 609.

40. Schreuder, G.M.T., et al., The HLA Dictionary 2004: a summary of HLA-A, -B, -C, -DRB1 /3/4/5 and -DQB1 alleles and their association with serologically defined HLA-A, -B, -C, -DR and -DQ antigens. Tissue Antigens, 2005. 65: p. 1 - 55.

41. Singh, P. B., R.E. Brown, and B. Roser, MHC antigens in urine as olfactory recognition cues. Nature, 1987. 327: p. 161 - 164.

42. Reynolds, J. B., B. S. Weir, and CC. Cockerham, Estimation of the coancestry coefficient: basis for a short-term genetic distance. Genetics, 1983. 105: p. 767 - 779.

43. Cavalli-Sforza, L. L. and A.W.F. Edwards, Phyligenetic analysis: models and estimation procedures. Evolution, 1967. 32: p. 550 - 570.

44. Nei, M., Genetic distance between populations. American Naturalist, 1972. 106: p. 283 - 292.

45. Midleton, D., et al., www.allelefreguencies.net New allele frequency database. Tissue Antigens, 2003. 61 : p. 403 - 407.

Claims

1. A test kit for the rapid determination of an individual's Major Histocompatibility Complex (MHC) profile, comprising: at least one solid substrate in contact with a proximal sample application zone, said solid substrate having N conjugation zones and N distal detection zones, said conjugation zones respectively containing N labelled primary binding reagents capable of binding with an analyte to form an analyte-primary binding reagent conjugate; said distal detection zone respectively having immobilized thereunto, N unlabeled secondary binding reagents capable of binding to a migrating analyte- labeled primary binding reagent conjugate to determine the presence or absence of N analytes which are indicative of defined N MHC antigens, wherein N is an integer from 6 to 30.

2. The test kit according to claim 1 , further comprising an item on which a unique identifier is recorded.

3. The test kit according to claim 1 or 2, wherein said N defined MHC antigens are N different MHC antigens selected from the group consisting of A^*01; A^*02; A^*0201; A^*03; A^*11 ; A^*1101 ; A*24; A*2402; B*07; B*0702; B^*08; B*0801; B*13; B^*1301 ; B*15; B^*1525; B^*35; B^*39; B*3901; B^*40; B*4001; B*4002; B^*44; B*48; B^*4801 ; Cw*01 ; Cw^*0102; Cw*03; Cw*0304; Cw*030401 ; Cw*04; Cw*0401 ; Cw*0403; Cw*0501 ; Cw*06; Cw^*0602; Cw^*07; Cw^*0701 ; Cw^*0702; Cw^*08; Cw^*0801 ; DPB1^*0201 ; DPB1^*0301 ; DPB1^*0401 ; DPB1^*0402; DPB1*0501; DQA1*0101 ; DQA1^*0102; DQA1^*0103; DQA1*0201; DQA1^*03; DQA1^*0301 ; DQA1*0401 ; DQA1*05; DQA1^*0501 ; DQB1^*02; DQB1^*0201 ; DQB1^*0202, DQB1*0301 ; DQB1*0302; DQB1*0402; DQB1*0501 ; DQB1^*06; DQBT0601 ; DQB1^*0602; DRB1*01 ; DRB1*03; DRB1^*0301 ; DRB1*04; DRB1^*07; DRB1^*0701 ; DRB1*08; DRB1*0802; DRB1*080302; DRB1*09; DRB1^*090102; DRB1^*11 ; DRB1^*110101 ; DRB1*12; DRB1*120201; DRB1^*13; DRB1*14; DRB1*1401 ; DRB1M5; DRB1*1501 ; DRB1*150101; and DRB1*160201.

4. The test kit according to claim 3, wherein said N different MHC antigens are selected from the group consisting of A*01, A^*02, A*0201 , A*11 , A^*1101 , A*24, A^*2402, B*1301, B*3901, B*4001, Cw^*03, Cw^*030401, Cw*07, Cw*0701, Cw^*0702, DPB1^*0201 , DPB1*0401 , DPB1^*0402, DQA1*0102, DQA1*03, DQA1*0301 , DQA1*05, DQA1^*0501 , DQBT02, DQB1*0201, DQB1*0301 , DQB1*06, DRB1*04, DRB1^*080302, and DRB1^*120201.

5. The test kit according to any one of claims 1 to 4, wherein said analytes are selected from the group consisting of MHC molecules, fragments of MHC molecules, naturally occurring derivatives of MHC molecules, MHC ligands, fragments of MHC ligands, naturally occurring derivatives of MHC ligands and combinations thereof.

6. The test kit according to any one of claims 1 to 5, wherein N is an integer from 8 to 12.

7. The test kit according to any one of claims 1 to 6, wherein said binding reagents are antibodies.

8. The test kit according to any one of claims 1 to 7, wherein each of said N unlabelled secondary binding reagents is permanently immobilized in one of said N detection zones respectively, and in each of N conjugation zones upstream from said N detection zones said N labelled primary binding reagents is mobile in said solid substrate in the presence of liquid.

9. The test kit according to any one of claims 1 to 8, wherein the label is selected from enzymes, fluorescent dyes, dye sols and gold sols.

10. The test kit according to any one of claims 1 to 9, wherein said at least one solid substrate is encased in a housing.

11. The test kit according to any one of the preceding claims, wherein said N defined MHC antigens are N different MHC antigens selected from the group consisting of A^*01 , A^*02, A^*0201, A*11 , A^*1101 , A^*24, A^*2402, B*1301 , B*3901 , B^*4001 , Cw^*03, Cw*030401 , Cw*07, Cw*0701 , Cw^*0702, DPB1*0201 , DPB1*0401 , DPB1^*0402, DQA1^*0102, DQA1*03, DQA1*0301, DQA1*05, DQA1^*0501, DQB1^*02, DQB1^*0201, DQB1^*0301 , DQB1*06, DRB1*04, DRB1 "080302, and DRB1*120201 ; and wherein said N primary binding reagents are N different antibodies capable of binding to said N different MHC antigens, respectively; and wherein said N secondary binding reagents are N different antibodies capable of binding to said N different MHC antigens, respectively.

12. A method of matching individuals, comprising the following steps: (a) obtaining a plurality of personal profiles from a plurality of individuals and storing said personal profiles in a database on a computer.

(d) storing said encrypted MHC profiles and the respective unique identifiers in a database on a computer, wherein each encrypted MHC profile and its unique identifier is allocated to one of said personal profiles;

(e) decrypting said encrypted MHC profiles using a key related to said unique identifier and a computer, and storing the decrypted MHC profiles in a database on a computer;

13. The method according to claim 12, wherein the quality of match between two individuals is defined as being high when more than 15% and less than 85% of the MHC antigens in the MHC profiles of the two individuals are the same.

14. The method according to claim 13, wherein the quality of match between two individuals is defined as being high when more than 25% and less than 75% of the MHC antigens in the MHC profiles of the two individuals are the same.

15. The method according to any one of claims 12 to 14, further comprising the step of providing to a first individual contact data of further individuals whose MHC profiles have a high quality of match with the MHC profile of said first individual.

16. The method according to any one of claims 12 to 15, wherein step (g) comprises generating a matrix containing qualities of match.

17. The method according to any one of claims 12 to 16, wherein steps (a), (b), (c) and (d) are carried out by a first service provider, and steps (e), (f) and (g) are carried out by a second service provider.

18. The method according to any one of claims 12 to 17, wherein the MHC profile of the individual consists of the presence or absence of at least five MHC antigens.

19. The method according to any one of claims 12 to 18, wherein said test kits provided in step (b) is a test kit according to any one of claims 1 to 10.

20. A method for the rapid determination of an encrypted MHC profile of an individual, in which a test kit according to any one of claims 1 to 10 is contacted via the sample application zone with a body liquid sample suspected of containing the analyte.

21. A method according to claim 20, wherein said body liquid sample is selected from human urine or human saliva or human sweat.

22. A method of determining the degree of match between a person wishing to adopt a child and a child, comprising using the test kit according to claim 1.